J Incl Phenom Macrocycl Chem
DOI 10.1007/s10847-017-0744-2
REVIEW ARTICLE
Essential oil–cyclodextrin complexes: an updated review
Geetika Wadhwa1 · Sunil Kumar1 · Lovely Chhabra2 · Sheefali Mahant3 ·
Rekha Rao1   
Received: 22 June 2017 / Accepted: 10 August 2017
© Springer Science+Business Media B.V. 2017
Abstract  Cyclodextrins, degradation product of carbohy-
drates, have been extensively exploited by food, pharmaceu-
tical and cosmetic industry by virtue of their ease of avail-
ability and their ability to entrap guest moieties. Various
cyclodextrin derivatives have been granted generally recog-
nized as safe (GRAS) status by several countries. The most
noteworthy characteristic of cyclodextrins is their ability to
form inclusion complexes with variety of molecules, impart-
ing protection and enabling solubility, bioavailability and
safety enhancement of challenging bioactives. In the last few
decades, investigations have revealed anti-microbial, anti-
inflammatory, insecticidal, analgesic and sedative properties
of essential oils. However, their poor solubility, volatility
and sensitivity to environmental factors pose challenge for
the formulation scientists. Inclusion complexes of essen-
tial oils with cyclodextrins have proved a useful strategy to
circumvent these challenges. The success of this approach
for essential oils is examplified by the commercial garlic
oil/β-cyclodextrin products, available under the trade names
Xund, Tegra, Allidex and Garlessence. Here, we present an
in-depth account of essential oil loaded cyclodextrin inclu-
sion complexes.
* Rekha Rao
rekhaline@gmail.com
1
Department of Pharmaceutical Sciences, Guru
Jambheshwar University of Science and Technology, Hisar,
Haryana 125001, India
2 nephrotoxicity limited its use in parenteral mode of deliv-
Department of Pharmaceutical Sciences, Maharishi
Markandeshwar University, Sadopur, Ambala 134007,
Haryana, India
3
Department of Pharmaceutical Sciences, Maharshi Dayanand
University, Rohtak, Haryana 124001, India
Keywords  Solubility enhancement · Kneading ·
Co-precipitation · Food · Cosmetic and pharmaceuticals
Introduction
Carbohydrates, in the form of starch and sucrose, are the
most common organic substances available in nature. From
very ancient times, they have been used for food and pro-
cessed through fermentation. The process of fermentation
results in mixture of mono-, di-, and oligo saccharides and,
under certain conditions, these degradation processes also
lead to the formation of cyclodextrins. The historical devel-
opment of these cyclic dextrins has been described in detail
by Loftsson and Duchene in their review [1].
Cyclodextrins are cyclic oligosaccharides with a hydro-
phobic cavity and hydrophilic surface. They are also ref-
ered to as cyclomaltose, cycloamylases and Schardinger
dextrins [2]. Cyclodextrins may contain a large number of
glucopyranose units (more than 15 per ring). Cyclodextrins
possess a truncated cone shape due to the presence of chair
form of glucopyranose units [3, 4]. Generally, they are cat-
egorized into naturally occurring and chemically modified
cyclodextrins [5]. The most common of naturally occur-
ring forms include α, β and γ cyclodextrins, consisting of
6, 7 and 8 glucopyranose units, linked by α-(1–4) bond [6].
Among these, β-cyclodextrin has been extensively used in
the early stages of pharmaceutical applications, owing to
its ease of availability and suitable cavity size for a wide
range of drugs. Although, its poor aqueous solubility and
ery [7]. With the view of extending the inclusion capacity
and the physicochemical properties, chemically modified
forms of cyclodextrin derivatives have also been prepared
and investigated. One of the derivatives of cyclodextrin,
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hydroxy propylated-β-cyclodextrin is regarded as a rela-
tively safer form, which is also non-irritating to the eye [8].
Derivatization can have a large influence on cyclodextrins
capacity to solubilize membrane components, as reported
by Motoyama et  al. [9]. Dimethylated-α-cyclodextrins
have been reported to release proteins form red blood cells
in rabbits. Some more modified forms of cyclodextrins
include, sulphobutylated-β-cyclodextrin, demethylated-
β-cyclodextrin, randomly methylated-β-cyclodextrin [4]
branched-β-cyclodextrins (maltosyl- and glucosyl-) and,
acetylated and sulphated cyclodextrins. Modifications of
cyclodextrins also yield alteration in solubility; for example,
methylated form of β-cyclodextrin deputes 50-fold higher
aqueous solubility than unsubstituted β-cyclodextrin [10,
11].
The most notable attribute of cyclodextrins is their ability
to create inclusion complexes with a wide variety of mol-
ecules (solid, liquid and gases) or their portions by molecu-
lar complexation [12]. Cyclodextrins act as host and a guest
moiety is entrapped within its cavity, resulting in complex
formation. During this inclusion phenomena, no covalent
bonds are formed or broken [13]. Only non-polar appropri-
ately sized molecules can be held in the lipophilic cavity
of cyclodextrin, producing inclusion complexes [14]. The
complexes can be formed, both, in solution or crystalline
form. During complex formation, the enthalpy-rich water
molecules are released from the cyclodextrin cavity and
guest moieties set up a dynamic equilibrium with cyclodex-
trin molecules in the inclusion complex [6].
Cyclodextrin-assisted encapsulation of guest molecule
offers numerous advantages namely solubility improvement
of poorly soluble molecules, enhancing stability of labile
moieties against degradation due to oxidation, light and
heat, control of sublimation and volatility, chromatographic
separation, physical separation of incompatible compounds,
masking unpleasant odors, taste modification and modified
release of flavors, fragrances and drugs. These advantages
open doors for the application of cyclodextrin complexes in
environmental protection [15], agriculture [16], biotechnol-
ogy [2, 17], packaging, textile industries [18], food [19],
pharmaceuticals [20] and cosmetics [21].
Essential oils, extracted from aromatic plants are vola-
tile compounds with strong odour. These secondary plant
metabolites are complex mixtures of many hydrocarbons,
terpenes, terpenoids and their derivatives. These oils, gener-
ally obtained by hydro or steam distillation or by a mechani-
cal process, usually consisting of 20–60 components. Out
of these, two or three are the main components, present at
reasonably greater concentration (20–70%) as compared to
others. It is the major constituents, which are responsible for
the biological properties of the essential oils, owing to the
presence of varying groups, which are derived from distinct
biosynthetic pathways [22]. Terpenoids are a class of natural
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J Incl Phenom Macrocycl Chem
compounds formed by C-5 building blocks, isopentenyl
diphosphate (IPP) and its isomer (DMAPP). Monoterpenes
(C-10), sesquiterpenes (C-15) and hemiterpenes (C-5) are
the representatives of this class. On the other hand, terpe-
nes carrying oxygen in the form of ether, hydroxyl, ketone,
aldehyde or carboxyl groups are termed as terpenoids [23].
Essential oils have found numerous medicinal uses in
humans as well as animals [24]. Externally, these are mostly
used in the form of gargles, mouthwashes and inhalations.
Though rarely used by oral route, these essential oils are
designated as safe and placed under GRAS category for
ingestion, where in these are administered after dilution
with olive oil, milk or soy milk. In addition, these oils are
generally considered safe for dermal application, where the
oil is present in diluted form in a formulation. However, such
an application can lead to skin reactions, especially, in case
of citrus oils which are sensitive to UV radiation and may
cause darkening of skin or irritation upon exposure to sun
[24]. When used in inhalation form, the concentrated vapors
of essential oils may lead to irritation in the eyes. It has been
found that essential oils undergo rapid metabolism, with
their distribution in the body being relatively high. Essen-
tial oils afford interesting applications in food, pharmaceuti-
cal field and cosmetics owing to their diverse and relavant
biological activities. However, due to their unstable nature
and fragile, and volatile constituents, they can undergo deg-
radation easily (by heat, light, oxidation, volatilization), if
they are not suitably protected from external environment.
Figure 1 represents the common challenges encountered
with essential oils.
In view of their properties relavant to human use, essen-
tial oils have been incorporated in several drug delivery
systems, carefully engineered to produce the desired effect
and overcoming their drawbacks at the same time. To name
a few, liposomes, micro and nanoemulsions, solid lipid
Fig. 1  The common challenges of essential oils
J Incl Phenom Macrocycl Chem
nanoparticles, have been developed [22]. Microencapsula-
tion of essential oils with β-cyclodextrins is another tech-
nique using the molecular inclusion method and helps in
improving the stability and efficacy of essential oils [25].
The chemical structures of active constituents of essential
oils complexed with cyclodextrins are depicted in the Fig. 2.
In order to further expand the applications of essential oils,
inclusion complexes have been widely explored by the
research fraternity in literature. The studies undertaken for
the entrapment of essential oils or their active moieties in
cyclodextrin complexes are reported in Tables 1, 2 and 3.
This review article presents an account of the recent
developments in the arena of essential oil–cyclodextrin com-
plexes. Preceding this, Marques et al., published an excellent
review on this topic in 2010 [65]. To the best of our knowl-
edge based on literature review, there is no updated review
publication in this area and our article may prove a timely
contribution for the research community. In this review,
the preparation techniques and characterization methods of
cyclodextrin complexes have been summarized. The advan-
tages and applications of cyclodextrin complexes entrapping
essential oils have also been discussed. This review can be
felicitous for scientists streaming in this area, to encapsulate
essential oils using cyclodextrin inclusion complexes. This
article also gives an insight into various factors influencing
inclusion complex formation, because an understanding of
the same is necessary for proper handling of these versatile
materials. However, our paper provides an overview of the
work done by various research groups in this arena, although
essential-oil loaded cyclodextrin complexes-based delivery
systems have been excluded, with the view to limit the scope
of the article.
Manufacturing methods for cyclodextrin inclusion
complexes
Apart from the ingredients incorporated in the formulation,
the preparation technique employed for manufacturing of
inclusion complexes plays an indispensible role, affecting
the product’s morphometrical properties as well as perfor-
mance. Literature reports a variety of preparation techniques
for encapsulating essential oils in cyclodextrin complexes.
Cyclodextrin complexes have been chiefly synthesized
using either co-precipitation method and kneading method
(Fig. 3).
In co-precipitation method, briefly, measured quanti-
ties of cyclodextrin and water are added to a flask placed
on magnetic stirrer, at a constant temperature (40/50 °C).
After stirring for an hour, saturated solution of cyclodex-
trin is obtained. Ethanolic solution of oil or active moiety
may be added dropwise to the saturated cyclodextrin solu-
tion, with continuous stirring for an hour, at the same rate
and temperature, as mentioned above. Then, this solution is
cooled at room temperature, with continuous agitation and,
maintained overnight at 4 °C. Finally, the cold, precipitated
inclusion complex can be recovered by vacuum filtration.
The filtrate is washed with suitable organic solvent, in order
to remove free guest components from the surface of cyclo-
dextrin and, finally they are freeze dried [28, 47, 48, 58].
In the kneading or paste method, accurately weighed
cyclodextrin is taken in porcelain motor. Adding small
amount of purified water, cyclodextrin is mixed with it
using pestle, resulting in the form of paste. Then, essential
oil or its components can be incorporated into the paste with
thorough mixing. The solid obtained should be washed with
small amount of suitable organic solvent and dried appropri-
ately, as mentioned in co-precipitation method [66].
Locci and coworkers prepared cyclodextrin inclusion
complexes loading carvacrol, thymol and eugenol using
super critical carbon dioxide. In this technique, measured
amount of cyclodextrin and guest molecule are loaded
into the autoclave thermostated and pressurized with car-
bon dioxide, at constant pressure and contact time. This is
followed by a rapid drop in pressure, which allows carbon
dioxide vaporization and separation of solvent free solid
inclusion complex. Although, poorly explored for inclusion
complexes of essential oils, this technique possesses great-
est advantage in terms of scale up for commercial applica-
tion, in comparison to classical methods using water, since
it leads to good yield of inclusion complex, an account of
higher solubility of guest molecules in carbon dioxide than
in water [56, 67].
In another technique, called molecular inclusion or self-
assembly aggregation, initially weighed quantity of cyclo-
dextrin is dissolved in specified amount of distilled water
and measured quantity of the guest is, then added to produce
a desired host:guest molar ratio. The mixture so obtained
is then placed in a shaking-incubator, maintained at appro-
priate rpm and temperature, for desired time period. The
obtained samples can be further dried to form solid inclusion
product [30].
In solvent evaporation technique, host and guest in spe-
cific ratio are dissolved in alcohol or some suitable solvent to
get a clear solution. The resulting preparation is allowed to
evaporate over night at room temperature. The cyclodextrin
complexes are, thus obtained [68].
In slurry method, slurry of weighed amount of
β-cyclodextrin is prepared in water. To this slurry, weighed
quantity of guest is added and stirred for 36 h by a magnetic
stirrer (400 rpm), the resultant product is filtered, and there-
after appropriately dried [57].
Conventionally, complexes are dried using hot air ovens,
vacuum ovens, spray dryers and freeze dryers. In case of
essential oils, most complexes should be dried below
40 °C to remove water. In addition, optimization of drying
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Fig. 2  The molecular structure of major components of essential oils investigated as inclusion complexes

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Fig. 2  (continued)

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Fig. 2  (continued)

temperature and other process conditions should also be
done, when highly volatile essential oils or their components
are taken as guests.
In literature, freeze drying method has also been
reported to have been used to prepare inclusion complexes
for essential oils. In these reports, solutions for cyclodex-
trin and guests were prepared in appropriate molar ratios
with agitation for 24 h and freeze dried after filtration [69].

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Table 1  Food and cosmetic applications of cyclodexrtin complexes with essential oils or their components
Sr. no. Active moiety Cyclodextrins Preparation technique Potential applications References

1 Lemon oil β-Cyclodextrin Precipitation method Flavor material [26, 27]

2 Mentha × villosa Hudson oil β-Cyclodextrin Kneading Used in cooking because of flavoring properties [28]
3 Cinnamon garlic β-Cyclodextrin Precipitation Food preservative (antifungal activity) [29]
4 Fish oil β-Cyclodextrin – Food manufacturing for mask the objectionable [30]
odour
5 Eugenol β-Cyclodextrin Molecular inclusion Flavor [31]
6 Cinnamon oil β-Cyclodextrin Precipitation method Condiment and flavoring material in food [32]
7 Linalool camphor Cyclodextrin – Food manufacturing [33]
8 Mentha piperita Cyclodextrin – Retention of aroma compound [34]
9 Linalool β-Cyclodextrin Slurry method Flavor carrier and food additives, decorative cos- [35]
metic, fine fragrance as in shampoos, soaps
10 Sweet orange β-Cyclodextrin Precipitation method Flavor [36]
11 Estragole Cyclodextrin – Food preservative and flavor [37]

Table 2  Pharmaceutical applications of cyclodextrin complexes with essential oils or their components

Sr. no. Active moiety Cyclodextrins Preparation technique Applications References

1 Lemon oil, garlic oil, allyl mus- β-Cyclodextrin – Volatility of oxidizable labile [38]
tard oil, marjoram oil, pepper- flavor substance
mint oil and tarragon oil
2 Lemon oil β-Cyclodextrin Precipitation Moisture, total oil, surface oil [39]
3 Starch and d-limonene β-Cyclodextrin – Stability enhancement of flavor [40]
4 Allium sativum α-β-Cyclodextrin Slurry method Thermal and oxidative stability [41]
5 Salvia sclarea β-Cyclodextrin – Stability enhancement [42]
6 Curcumin HPβ-cyclodextrin Solvent evaporation Improved solubility [43]
Mβ-cyclodextrin
7 Litsea cubeba β-Cyclodextrin Suspension method Stabilization of essential oil [44]
8 Eugenol HP-β-Cyclodextrin Precipitation Improved solubility [45]
9 Eugenol β-Cyclodextrin Self-aggregation Release characteristics and stor- [46]
age temperature
10 β-Caryophyllene β-Cyclodextrin Co-precipitation Increased oral bioavailability [47]
11 Xiang–Fu–Si–Wu β-Cyclodextrin Co-precipitation Increased oral bioavailability [48]
12 Catfish β-Cyclodextrin Kneading and co-precipitation Mask objectionable odour [49]
13 Folium artemisia argyi β-Cyclodextrin – Stabilization of essential oil [50]

Factors affecting complexation of essential oils
with cyclodextrins
In order to fabricate cyclodextrin inclusion complexes for
essential oils or their active constituents, with desirable
characteristics, it is important to be well versed with the in
depth knowledge regarding various factors that may influ-
ence process of complex formation. Factors playing vital
role in cyclodextrin complexes have been discussed below.
Type of cyclodextrins
Type of cyclodextrin may affect encapsulation of essen-
tial oils in cyclodextrin inclusion complexes. Amongst the
various forms of cyclodextrins, β-form is the most explored
by various research groups, owing to its ability of taste
masking, improving solubility and ensuring stability. For
encapsulation of Salvia sclarea essential oil (SEO), vari-
ous cyclodextrins like β-cyclodextrin (β-CD), DM-β-CD
[(2,6-dimethyl)-β-cyclodextrin], 2-HE-β-CD [mono{2-O-(2-
hydroxyethyl)}-β-cyclodextrin] and 2-HP-β-CD [mono{2-O-
(2-hydroxypropyl)}-β-cyclodextrin] were investigated and,
among these, β-CD showed stronger inclusion interactions
with SEO [42]. Recently, in a similar study, β-cyclodextrin
DM-β-CD and HP-β-CD were investigated for encapsulation
of Folium artemisia argyi essential oil (FAA EO). Fluores-
cence spectrometry was used to study inclusion interaction
for β-cyclodextrin derivatives [50]. This parameter needs to

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Table 3  Therapeutic applications of cyclodextrin complexes with essential oils or their major components
Sr. no. Active moiety Cyclodextrins Preparation technique Applications References

1 Sardinian thymus herba – – Antibacterial [51]

2 Curcumin HPβ-cyclodextrin Solvent evaporation Antiangiogenic antiinflammatory [43]
Mβ-cyclodextrin
3 Thyme β-Cyclodextrin Precipitation method Antifungal [52]
4 Citronella oil, citronellal and citron- β-Cyclodextrin Kneading method Mosquito repellent [53]
ellol
5 Geraniol β-Cyclodextrin Slurry method Anticancer antimicrobial insect repel- [54]
lent
6 Linalool β-Cyclodextrin – Antinociceptive effect [55]
7 Lycopene β-Cyclodextrin – Super critical precipitation [56]
8 Lippia grata β-Cyclodextrin Slurry method Pain modulation [57]
9 Satureja montana β-Cyclodextrin Co-precipitation Antifungal activity antioxidant [58]
10 Thymol and thyme oil β-Cyclodextrin Kneading method Antimicrobial [59]
11 Coriander β-Cyclodextrin Co-precipitation Antioxidant antimicrobial [60]
12 Carvacrol β-Cyclodextrin Agitation Improve cancer pain [61]
13 Thymus catharinae camarda β-Cyclodextrin – Antimicrobial [62]
14 Linalool β-Cyclodextrin – Gastric lesion [63]
15 Citronellal β-Cyclodextrin Co-evaporation Anti hyperalgesic [64]

Fig. 3  Fabrication of cyclodextrin complexes using co-precipitation technique

be further explored in furturistic studies involving essential
oil cyclodextrin complexes.
Host–guest ratio
Ratio of guest and host can affect inclusion efficiency, an
important characteristic feature of inclusion complexes. In
addition, for essential oils, maximum retention of volatiles
plays a major role and needs to be optimized in prepared
inclusion complexes. Most of the literature on essential
oil inclusion complexes reports investigations using single
ratio of essential oil/essential components and cyclodextrins.
Either guest or host amount can be varied for the preparation
of cyclodextrin complexes, or their ratio may be optimized
(Table 4).
Songkro et  al. prepared-cyclodextrin complexes by
kneading technique using two weight ratios: 1:1 and 1:2,
of citronella oil and β-cyclodextrin. It was observed that

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Table 4  Guest host ratio along with preparation techniques of cyclodextrin complex with essential oil or their constituents
Sr. no. Essential oil or their con- Preparation technique Guest host ratio In vivo activity References
stituents

1 Thymol and carvacrol Co-precipitation method – – [70]

2 Lemon oil Precipitation method 3:97, 6:94, 9:91, 12:88, Effect on the inclusion [26]
15:85 efficiency
3 Lemon oil Kneading – – [27]
4 Lemon oil Precipitation – – [39]
5 Sardinian Thymus herba- – – – [51]
barona Loisel
6 Carvacrol, thymol and Supercritical carbon dioxide 1:25 – [67]
eugenol technique
7 d-Limonene Kneading and slurry method – Flavor retention [40]
8 Allium sativum – – – [41]
9 Menthe x villosa Hudson oil Co-precipitation and knead- – – [28]
ing
10 Salvia sclarea Shaking method 1:1 Inclusion interaction [42]
11 Cinnamon leaf and garlic oil Precipitation 0:100, 4:96, 8:92, 12:88, – [29]
16:84
12 Litsea cubeba Precipitation 1:1 – [44]
13 Thyme Precipitation method 0:100, 4:96, 8:92, 12:88, Antifungal [52]
14:86
14 Eugenol – 1:1 – [45]
15 Cinnamon oil Precipitation 5:95, 10:90, 15:85, 20:80 – [32]
16 Eugenol Freeze drying 1:1 – [31]
17 Geraniol Paste and slurry method – – [54]
18 Linalool and camphor – – – [33]
19 Citronella oil, citronellal and Kneading method 1:1, 1:2 Mosquito repellent [53]
citronellol
20 Eugenol Self-aggregation or molecu- 1:1 – [46]
lar inclusion
21 Caryophyllene Co-precipitation – – [47]
22 (−) Linalool – 1:1 Antinociceptive [55]
23 Menthe piperita – – – [34]
24 Thymol and thyme oil Kneading method and freeze 1:1 Antimicrobial [59]
drying
25 Sweet orange flavor Precipitation – – [36]
26 (−)-Linalool Paste and slurry method 1:1 – [35]
27 Coriander Co-precipitation 5:95, 10:90, 15:85, 20:80 Antioxidant Antimicrobial [60]
28 Satureja montana Co-precipitation 5:95, 10:90, 15:85, 20:80 Antifungal Antioxidant [58]
29 Lippia grata Slurry process 1:1 Reduce orofacial nociception [57]
in mice
30 Xiang–Fu–Si–Wu Co-precipitation method – Inc. oral bioavailability in [48]
rats
31 Estragole Freeze dried method 1:1, 1:3, 1:6 – [37]
32 Carvacrol Slurry method – – [61]
33 (−)-Linalool – – – [63]
34 Citronellal Co-evaporation – – [64]
35 Clarias batrachus oil Co-precipitation and knead- 1:8 – [49]
ing
36 Thymus catharine Camarda – – Antimicrobial activity [62]
37 Folium artemisia argyi – – – [50]

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different ratios resulted in complexes of different color: yel- [29]. From the findings of these studies, it may be concluded
low (weight ratio 1:1) and white (weight ratio 1:2), due to that saturation of cyclodextrin reaches for guest at a particu-
difference in the amount of β-cyclodextrin used. From scan- lar ratio and, therefore optimization of the process should be
ning electron microscopy (SEM) analysis, increase in size in established with respect to the highest recovery of essential
agglomerated inclusion complexes was seen, on increasing oil volatiles.
weight ratios from (1:1 to 1:2). In Fourier transform infra red
spectroscopy (FTIR) analysis, lower shifts were noted with
Preparation techniques
1:1 (citronellal and β-cyclodextrin) weight ratio, but higher
shifts were obtained with increase in weight ratio up to 1:2.
As discussed in “Manufacturing methods for cyclodextrin
From differential scanning calorimetric (DSC) analysis, the
inclusion complexes” section, the inclusion complexes can
formation of complex was clearly confirmed for the ratio
be prepared using different techniques. Selection of prepa-
1:2 due to absence of citronellal peak, unlike 1:1 complex.
ration method directly affects inclusion efficiency of essen-
Based on mosquito repellent action, citronella oil inclusion
tial oils with cyclodextrins (Table 4). Martins et al. used
complex containing lotion (weight ratio 1:1) demonstrated
co-precipitation and kneading technique for encapsulating
promising results, along with controlled release behavior, as
Mentha × Villosa Hudson oil in β-cyclodextrin cavity. The
compared to plain citronella oil [53].
co-precipitation method showed better results since, in this
All other researchers who investigated variable ratios
method the recovery of oil from complex powder was 95.9%,
used co-precipitation method and, investigated their
resulting in lesser material loss [28]. Tao and coworkers
effect on inclusion efficiency and retention of essential
synthesized thymol and thyme oil β-cyclodextrin inclu-
oil. Bhandari and coworkers explored, five lemon oil to
sion complexes by freeze drying and kneading method. The
β-cyclodextrin ratios (3:97, 6:94, 9:91, 12:88, 15:85 w/w),
entrapment efficiency of β-cyclodextrin inclusion complexes
in order to determine their effect on the inclusion efficiency.
prepared by freeze drying was found to be higher than those
Retention of lemon oil volatiles reached maximum at the
synthesized by kneading, for both thyme oil and thymol. In
lemon oil to β-cyclodextrin ratio of 6:94. However, maxi-
kneading method, complexation and drying are carried out
mum inclusion capacity was obtained at ratio 12:88, in
in an open container at room temperature, resulting in loss of
which complex contained 9.68% (w/w) lemon oil [26]. In
essential oil, owing to volatilization and difference in entrap-
a similar study, Petrovic and his research group fabricated
ment values. Results of antimicrobial activities of prepared
inclusion complexes using co-precipitation technique, with
complexes carried out in pork meat system revealed that
four ratios of cinnamon oil to β-cyclodextrin (5:95, 10:90,
thymol and thyme oil β-cyclodextrin complexes fabricated
15:85 and 20:80 w/w). The formulation containing 15:85
by freeze drying possessed greater (p < 0.05) anti-microbial
w/w (oil:β-cyclodextrin) exhibited maximum inclusion effi-
activity, when compared to MIC values of free thymol and
ciency (117.2 mg oil/g of β-cyclodextrin), whereas the for-
thyme oil, whereas, complexes prepared by kneading method
mulation with ratio 10:90 (oil to β-cyclodextrin) displayed
did not represent such enhancement [59]. In the same year,
maximum retention of essential oil volatiles (94.18%) [32].
Menezes et al. fabricated (−)-linalool β-cyclodextrin com-
Haloci and co-workers investigated four ratios of Satureja
plexes using paste and slurry methods. Results indicated that
montana essential oil, (5:95,10:90, 15:85 and 20:80) and
total (−)-linalool entrpped in complex cavity was 8.6% by
maximum retention of essential oil volatiles reached upto
paste method while by slurry method 42.57% (−)-linalool
93.15% (at oil to β-cyclodextrin ratio15:85) and maximum
was found to have been entrapped in complex cavity [35].
inclusion efficiency was achieved at ratio 20:80 w/w (oil
to β-cyclodextrin) [58]. However, in the study by Dima
et al. both maximum inclusion efficiency and maximum Effect of drying
retention of oil (94.23%) was attained with ratio 15: 85
(oil and β-cyclodextrin) [60]. Del Toro-sanchez et al., var- Bhandari et  al., reported fiffteen min of kneading time
ied thyme oil concentration and fixed the concentration of as the optimum period for encapsulation of lemon oil in
β-cyclodextrin and in their study five ratios were investi- β-cyclodextrin. This group further reported that the fla-
gated (0:100, 4:96, 8:92, 12:88 and 14:86). From the results vor inclusion profile in the cyclodextrin complex does not
of recovery of essential oil from inclusion complex, it was depend on the kneading time. In this study, kneading was
found that 8:92 ratio (oil to β-CD) had highest thymol con- followed by drying of paste using, two modes: spray dry-
tent. These results were also confirmed by gas chromatogra- ing and vacuum drying and, the results were compared.
phy mass spectrometry (GC–MS) data [52]. The saturation Both drying methods were found similar, when extent of
patterns of garlic oil in β-cyclodextrin inclusion complexes encapsulated oil was measured. However, spray drying was
were observed at 12:88 ratio (oil:β-cyclodextrin), whereas described as a simpler and quicker method in comparison to
for cinnamon oil, no saturation was found up to 14:86 ratio vacuum drying [27]. Further, due to its efficiency at lower

13
J Incl Phenom Macrocycl Chem

drying temperature, spray drying can prove more efficient Characterization aspects of essential oil–
for encapsulating volatile oils in cyclodextrin complexes. cyclodextrin complexes

To study the interaction of cyclodextrins and loaded guest

moiety and, to understand the process of their design and
fabrication, various analytical techniques are used. The
cyclodextrin complexes are characterized by investigating
the following studies (Table 5).

Table 5  Characterization of essential oil and their components loaded in cyclodextrin complexes

Sr. no. Drug/oil Characterization techniques References
1
1 Thymol and carvacrol H-NMR [70]
2 Lemon oil Moisture determination, capillary GC–MS [26]
3 Lemon oil Moisture detemination [27]
4 Lemon oil GC, capillary GC–MS [39]
5 Sardinian Thymus herba-barona Loisel GC, GC–MS, 13C-NMR [51]
13
6 Carvacrol, thymol and eugenol C-CPMAS, 1H-NMR [67]
7 Lippia sidoides oil TGA, XRD, mass spectroscopy [66]
8 d-Limonene Water absorption index,water solubility index, specific mechani- [40]
cal energy
9 Allium sativum DSC, SEM, TGA, GC–MS [41]
10 Menthe x villosa Hudson oil GC–MS, thermal analysis, XRD [28]
11 Cinnamon leaf and garlic oil GC–MS, moisture determination, IR [29]
12 Litsea cubeba GC–MS, UV, FTIR [44]
13 Fish oil GC–MS, AFM, particle size and zeta potential [30]
14 Eugenol DSC, water content measurements, thermal properties, particle [31]
size and zeta potential
15 Thyme GC–MS, IR, 1H-NMR [52]
16 Eugenol Phase solubility studies, FTIR, NMR [45]
17 Cinnamon oil Moisture determination, GC–MS [32]
18 Citronella oil, citronellal and citronellol SEM, FTIR, DSC [53]
19 Eugenol Particle size and zeta potential analysis, TEM [46]
20 Geraniol Moisture determination, thermal analysis, FTIR, SEM [54]
21 Linalool and camphor GC–MS, molecular modeling [33]
22 (−)-Linalool – [55]
23 Caryophyllene UV–Visible spectrophotometry, FTIR, DSC, GC–MS [47]
24 Menthe piperita GC–MS, molecular modeling [34]
25 Coriander – [60]
26 Satureja montana GC–FID [58]
27 Thymol and thyme oil Particle size, phase solubility, DSC, TEM [59]
28 Lippia grata GC–MS, DSC, thermal analysis, X-ray diffraction, SEM [57]
29 Sweet orange flavor Thermogravimetric analysis [36]
30 (−)-Linalool Moisture determination, DSC, XRD, FTIR, GC–MS [35]
31 Xiang–Fu–Si–Wu Decoction essential oil DSC, FTIR, XRD, SEM, LC-MS [48]
32 Carvacrol Thermal analysis, SEM [61]
33 Estragole UV–Visible spectrophotometry, GC, NMR, DSC, FTIR [37]
34 (−)-Linalool – [63]
35 Citronellal Thermal analysis, moisture determination, FTIR, SEM [64]
36 Cat fish oil SEM, FTIR, DSC [49]
37 Folium artemisia argyi GC–MS, FTIR [50]
38 Thymus catharine Camarda GC–MS, NMR [62]

13

Phase solubility studies
In order to investigate the solubility limit, phase solubil-
ity studies can be performed. These studies are helpful in
determining phase solubility constant and stability con-
stants. Generally, these studies are performed according
to the method given by Higuchi and Connors, 1965 [59].
In order to explore molecular association of cyclodextrin
with guest, an excess amount of guest moiety can be added
to varying concentration of aqueous cyclodextrin solution.
The suspension is shaken in screw cap vials using mag-
netic stirrer for a time period until equilibrium is attained.
The samples are centrifuged at 500 rpm to separate insolu-
ble oil/oil components and filtered through a syringe filter.
The filtered solution is appropriately diluted and analyzed
spectroscopically. Phase solubility diagram can be plotted
with guest solubility as a function of cyclodextrin concen-
tration, according to equation given below.
slope
K= (1)
SO(1 − slope)
where SO is the aqueous solubility of oil or intercept of
plot (in mol/l) and K is the stability constant. Initially, the
solubility of host increases linearly with concentration of
cyclodextrin. At one stage, increasing the concentration of
cyclodextrin does not enhance solubility of guest, signifying
the solubility limits within the cyclodextrin concentration
range. The phase solubility profile data alone do not con-
firm formation of inclusion complexes. These only explain
how drug solubility is influenced by increasing cyclodextrin
concentration [71].
The value of K from Eq.  (1), between 200 and
5000 m −1, is considered most suitable for the enhance-
ment of stability and solubility of poorly soluble drugs
[43]. Additionally, phase solubility experiments can be
performed to determine thermodynamic properties of the
cyclodextrin complex formed. Gibbs free energy, a ther-
modynamic function can also be calculated. Complexa-
tion lowers the thermodynamic potential of the dissolved
moiety [65].
The driving forces for cyclodextrin complex formation
include Vander-Wall interaction, electrostatic interaction,
hydrophobic interaction, hydrogen bonding, charge-trans-
fer interaction and release of enthalpy rich water moieties
from the cyclodextrin cavity [14, 72].
Although, phase solubility is an important study
for cyclodextrin complexes, only a limited number of
researchers have performed it for essential oils or their
constituents. Such studies have been carried out for pure
α-bisabolol and chamomile essential oils [73], eugenol
[45] and thymol and thyme oil [59].
13
J Incl Phenom Macrocycl Chem
Fourier transform infra‑red spectroscopy
Fourier transform Infra-red spectroscopy is the major
technique to determine the presence of functional groups.
Formation of intra-molecular hydrogen bond/complex for-
mation between the cyclodextrin and host moieties can be
confirmed with the aid of this technique. The appearance of
new resultant peaks in the FTIR spectrum indicates bond
formation between host and guest. Further, due to included
part of guest molecule, IR bands are either shifted or their
intensities altered. FTIR spectra of the pure oil/oil constitu-
ents, cyclodextrin, physical mixture and guest-loaded cyclo-
dextrin complex can be determined and compared.
Songkro et al. carried out FTIR analysis of citronella oil,
physical mixture of citronella oil and β-cyclodextrin and
inclusion complex of citronella oil. This group observed
both physical mixture and inclusion complex weaken some
peaks of pure citronella oil such as 1016 and 1617 cm−1. A
higher O–H bonding (1645 cm−1) shift was found for physi-
cal mixture, 1:1 and 1:2 inclusion complexes. In inclusion
complexes, changes in O–H stretching bands for citronella
oil (3413 cm−1) were demonstrated, suggesting the interac-
tion between citronella oil and β-cyclodextrin. This group
also noted that in 1:1 inclusion complex lower shifts were
observed, while higher shifts were seen with 1:2 (citronella
oil and β-cyclodextrin inclusion complex) [53].
In 2015, Xi et al. prepared Xiang–Fu–Si–Wu Decoc-
tion essential oil (XFSWD) β-cyclodextrin complex. This
group has presented FTIR spectra of pure β-cyclodextrin,
physical mixture and inclusion complex and reported dis-
appearance or reduction of peak intensities, on formation
of inclusion complex [48]. In the following year, Zi-Tao
Jing et al. prepared molecular microcapsules of F. arte-
misia Argyi (FAAEO) using β-cyclodextrin derivatives.
This group carried out FTIR analysis of essential oil alone,
β-cyclodextrin alone, glucose-β-cyclodextrin, physical mix-
ture and prepared microcapsules. For comparison purpose,
the major ratios of FAAEO and β-cyclodextrin were kept
constant in physical mixture as well as microcapsules. When
all FTIR spectra were compared, it was observed spectra
of β-cyclodextrin exhibited major peaks at 3391  cm −1
(OH), 2924 cm−1 (CH), 1649 cm−1 (=O), 1156 cm−1 (–O)
and 1028 cm−1 (C–O–C). Very strong absorbance peak at
1739  cm−1 was seen for the C=O (stretching) vibration
of FAAEO. The spectrum of physical mixture depicted
the peaks of both FAAEO and β-cyclodextrin, but with
decreased peak intensity. Additionally, a number of peaks
of FAAEO at 3084, 2962, 2725, 1096 and 986 cm−1 were
found disappeared representing strong physical interaction
of essential oil with β-cyclodextrin in microcapsules, the
peaks of FAAEO at 3084, 2962, 1096 and 986 cm−1 were
found disappeared as in physical mixture and the peaks of
essential oil at 3459, 2930 and 1739 cm−1 were decreased
J Incl Phenom Macrocycl Chem
in intensity showing that (C=O stretching) vibration was
restricted after formation of the inclusion complex. The
C=O with carboxylic functional group of the guest essential
oil had been encapsulated in the hydrophobic cavity of the
host. Hence, the FTIR analysis confirmed the formation of
microcapsules of FAAEO to β-cyclodextrin [50].
Such studies have also been reported for cinnamon leaf
and garlic oil [29], Litsea cubeba [44], thyme [52], catfish
(Clarias batrachus) oil [49] and essential oil components
like geraniol [54] and citronellal [64].
Thermal analysis
Differential scanning calorimetry
Another technique reported to clarify the interaction
between guest and host is differential scanning calorimetry
(DSC). It is generally observed that the cyclodextrin cavity
affects the thermal behavior of inclusion complexes, thus,
leading to shift in their endothermic peaks as compared to
pure compounds. Shifting, broadening and appearance of
new peaks or disappearance of certain peaks in the host may
be due to evaporation, oxidation, decomposition, melting
or polymorphic transition, suggesting complex formation.
Inclusion complex formation generally causes reduction or
absence of endothermic peaks of host (at the temperature of
its boiling and melting point) [74].
Seo and his research group carried out DSC for eugenol,
β-cyclodextrin, their physical mixture and eugenol-loaded
inclusion complex. The thermogram of β-CD–eugenol
complex showed thermal transitions at 196.8 °C with endo-
thermic peaks. Pure β-CD peaks appear at 187.3–188.7 °C.
This may be due to host and guest interaction culminating
in increase phase transition temperature. Further, peak cor-
responding to boiling point of eugenol was found to have
disappeared, pointing towards inclusion complex forma-
tion between β-CD and eugenol. Partial inclusion reac-
tion was also observed in eugenol physical mixture due to
similar transition temperature of eugenol–β-CD inclusion
complexes [31]. Similar studies were performed for Allium
sativum [41], citronella oil, citronellal and citronellol [53,
64], geraniol [54], β-caryophyllene [47], thymol and thyme
oil [59], Xiang–Fu–Si–Wu Decoction essential oil [48],
estragole [37] and catfish (C. batrachus) oil [49].
Thermogravimetric analysis
Thermogravimetric analysis (TGA) is employed to under-
stand the thermostability, melting point and crystalline
behavior of particles. Data from TGA may be used for
confirmation of DSC results [75]. Changes in weight loss
provide a supporting evidence for the formation of inclu-
sion complexes. Menenzes and his coworkers analyzed
geraniol–β-CD complexes using TGA. Thermogravimetry/
differential thermo gravimetry (TG/DTG) curves of the
geraniol, β-CD, their physical mixture and inclusion com-
plexes (prepared by paste and slurry method) were analyzed.
The mass losses calculated by their data analysis from spe-
cific intervals for each one have been reported. Percentage
of geraniol evaporated was found upto 230 °C, mass loss due
to evaporation of geraniol and water release up to 120 °C
from physical mixture and inclusion complex. In addition,
percentage of water released from β-CD upto 120 °C was
obtained. Further, mass loss was also observed from physical
mixture and geraniol inclusion complex, which was attrib-
uted to geraniol release (125–270 °C) and thermal decom-
position of β-CD, physical mixture and inclusion complex
(270–391 °C). Finally, elemental carbon formation from
β-CD, physical mixture and geraniol inclusion complexes
occurred due to sample carbonization from 365 to 900 °C
[54]. Inclusion complexes of A. sativum [41], geraniol [54]
and β-caryophyllene [47] have been characterized by TGA
as per the literature.
X‑ray diffraction analysis
Another simple and useful method for evaluating chemical
decomposition and complex formation is powder-ray dif-
fractometry. X-ray diffraction (XRD) pattern of the sam-
ple is usually determined as a function of scattering angle
[76]. As a result of complex formation, shifting of certain
peaks, sharpening of the existing peaks and disappearance
or appearance of a few new peaks occurs. This charac-
terization technique is the most beneficial method for the
confirmation of inclusion complexes, either essential oils
or volatile components, as liquid guest molecules show no
diffraction patterns and differences in the diffractograms of
uncomplexed cyclodextrin and inclusion complex can be
easily established [54, 77, 78]. Menezes and his research
group investigated β-cyclodextrin, physical mixture and
β-cyclodextrin inclusion complexes (prepared by paste and
slurry method) of (−)-linalool by XRD. β-cyclodextrin was
found to be crystalline in nature. It was observed that inclu-
sion complex formation led to large shift in the XRD signals
of β-cyclodextrin. Also, inclusion complexes represented
different XRD signals in comparison to β-cyclodextrin and
physical mixture. Inclusion complexes further, showed weak
interaction, demonstrating the formation of inclusion com-
plex between (−) linalool and β-cyclodextrin [35].
However, review of the published literature reveals that
this technique is poorly explored for essential oils and their
constituents. This method has been reported for Mentha ×
villosa Hudson oil [28], linalool [35], Lippia grata leaf [57],
Xiang–Fu–Si–Wu Decoction essential oil [48] and estragole
[37].
13

Microscopic techniques
Microscopic techniques like SEM and transmission elec-
tron microscopy (TEM) can be employed for evaluation
of particle size and shape and to carry out morphological
analysis of cyclodextrin complexes [79]. Another micro-
scopic technique, atomic force microscopy (AFM) may
give the information regarding topography of inclusion
complexes, complementing well established analytical
techniques like X-ray crystallography, nuclear magnetic
resonance (NMR), and, light and electron microscopy.
AFM, a modern technique, helps to minimise the basic
drawbacks of scanning microscopy (only image conduct-
ing or semiconducting surfaces).
SEM analysis is useful for visualizing the surface
texture of the preparations [80]. In SEM, the developed
particle is imparted conductivity under vacuum, using
focused electron beam. Siqueira-Lima et al. encapsulated
Lippia grata leaf essential oil using β-cyclodextrin. From
SEM results, this group reported that there were drastic
changes in the original morphology and particle shape of
β-cyclodextrin, in inclusion complex. On the other hand,
particle morphology and shape of the corresponding phys-
ical mixture were found to be similar to β-cyclodextrin
[57]. Further, essential oil and β-cyclodextrin complexes
appear as agglomerates in microscopic analysis [81].
This technique has been used to study A. sativum [41],
Xiang–Fu–Si–Wu decoction [48], thymol and thyme oil
[59], carvacrol constituent of Oregano [61], citronellal
constituent of Cymbopogon species [64] and citronella oil,
citronellal and citronellol [53], eugenol [46] and geraniol
[54] cyclodextrin complexes.
Transmission electron microscopy can be used for inves-
tigating morphology of particles suspended in liquids. Mor-
phological analysis of TEM have been reported for euge-
nol [31], thymol and thyme oil [59]. Seo and his research
group observed large aggregates of eugenol–β-cyclodextrin
complex as an agglomerated form which resulted from self-
assembly of cyclodextrins in water [31].
Moisture content determination
Studies for the determination of moisture in cyclodextrin
complexes can be performed by Karl Fisher method, vac-
uum oven drying method or toluene distillation. Moisture
determination helps in supporting TG analysis, as TG cannot
differentiate between water and oil mass losses from inclu-
sion complexes [82]. Hence, moisture determination can be
used for estimating total volatile constituents loss from TG
curves. It has been carried out for lemon oil [26], cinnamon
leaf and garlic oil [29], cinnamon oil [32], geraniol [54] and
linalool [35] cyclodextrin inclusion complexes.
13
J Incl Phenom Macrocycl Chem
Zeta potential and polydispersity index
Stability of the formed inclusion complexes can be estimated
by zeta potential assessment. Generally, at lower zeta poten-
tial values, attractive force between particles exceed repul-
sive forces, leading to aggregation or coalescence of parti-
cles [83]. Zeta potential is a measurement of surface charge
that affects interaction of formulation with the biological
environment. Zeta potential can be estimated by measuring
electric potential, i.e., electrophoretic mobility and diffu-
sion coefficient [81]. The pH and electrolyte concentration
should be considered while measuring zeta potential [84,
85]. Seo et al. determined zeta potential of eugenol–cyclo-
dextrin complex using Zeta sizer. It was found to be highly
stable with zeta potential and PDI of −34.5 ± 1.2 mv (aver-
age value) and 0.3, respectively [31].
Polydispersity index (PDI) is a measure of uniform distri-
bution of particle size present in the cyclodextrin complex
suspensions, reflecting its tendency to agglomerate [86]. As
a result of the self-assembly of cyclodextrin in water, there
is strong tendency of cyclodextrin inclusion complexes for
agglomeration [31]. Hence, PDI measurements can prove as
a useful tool for stability analysis of cyclodextrin complexes.
Nuclear magnetic resonance spectroscopy
Nuclear magnetic resonance (NMR) spectroscopy may pro-
vide direct evidence for the entrapment of guest into the
cyclodextrin cavity. This technique has been used to verify
inclusion of numerous essential oils, or their constituents:
thymol and carvacrol [70], eugenol [45], estragole [37]
and Thymus catharinae camarda [62]. In cyclodextrins,
generally, shift for hydrogen atoms of cyclodextrin in the
cyclodextrin guest complex with respect to free cyclodex-
trin can be measured [67, 70]. If a guest moiety is encap-
sulated into cyclodextrin cavity, the H-atoms (C–3H) and
(C–5H) located in the interior of the cavity will be signifi-
cantly shielded by the guest and exhibit a considerable up
field shift [87, 88]. Whereas, H-atoms (C–2H, C–4H and
C–6H) on the outer surface will show a marginal up field
shift [89]. H-atoms of the guest moiety depict correspond-
ing downward shift characterized β-cyclodextrin carvacrol
inclusion complex using 1H-NMR spectroscopy. Remarkable
high yield shift for H3–H5 β-cyclodextrin proton resonance
were measured in β-cyclodextrin–carvacrol inclusion com-
plex, which indicated the true formation of this inclusion
complex. The inclusion of carvacrol in the hydrophobic cav-
ity of β-cyclodextrin includes this shielding of H3 or H5.
Moreover, a strong interaction between the H6 proton (in rim
of β-cyclodextrin) and guest was also reported [62].
C13 NMR has been applied for Sardinian Thymus
herba-barona Loisel [51], carvacrol, thymol and eugenol
[67], inclusion complexes. Using this technique, interaction
J Incl Phenom Macrocycl Chem
between the cyclodextrin cavity and guest moiety can be
studied from the shifting of carbon atoms.
Spectroscopic techniques
Sometimes, a bathochromic shift or/and band broadening
takes place on complex formation leading to alteration in
the original absorption spectrum (visible or ultra-violet) of
the guest moiety. This shift may be due to a partial shield-
ing of the excitable electrons in the cyclodextrin cavity [90].
High electron density in the cyclodextrin cavity may result in
mobilization of electrons of the entrapped guest moiety [91].
This phenomenon can be explored for essential oil loaded
cyclodextrin complexes.
Raman spectroscopy may prove useful in revealing
structural features which are not easily accessible by others
spectroscopic techniques. Although, most of the essential oil
cyclodextrin complex studies provide IR data as an evidence
for inclusion complex formation, but Raman spectroscopy
may provide clearer and better evidence of formation of
inclusion complex. This technique has been used for valida-
tion in curcumin cyclodextrin (1500–1800 cm−1) complex
and revealed their complex formation [92]. Hence, this tech-
nique can be explored for characterization of essential oil
cyclodextrin complexes.
Chromatographic techniques
Although an old technique, thin layer chromatography finds
a lot of application in pharmaceutical analysis. It plays a
vital role in the early stages for specific quantitative analysis
using spot illusion followed by spectrophotometric detec-
tion. Another chromatographic technique, gas chromatogra-
phy is a crucial separation technique for detection of volatile
compounds, therefore, it has been widely used for essential
oils. It allows accurate quantitative determination due to
combination of separation and online detection [93]. Some
groups of authors used this technique for analysis of lemon
oil [39], Sardinian T. herba-barona Loisel [51], estragole
[37] and cyclodextrin complexes.
Gas chromatography is conjugated with mass-spec-
trometry for quantitative analysis of complex mixtures. In
case of essential oils, the GCMS profile helps in identifica-
tion of major components along with quantities present in
oils. Therefore, it plays a vital tool to study composition of
essential oil and investigating their bioactivities along with
mechanism of action. For essential oil inclusion complexes,
this technique have been used for analysis of lemon oil [26],
Mentha villosa [28], Litsea cubeba [44], cinnamon oil [32],
thyme [52], Mentha piperita [34], (−)-linalool [35], Lippia
grata [57] and F. artemisia [50].
Another sophisticated technique, liquid chromatography
mass spectroscopy (LCMS) has also been reported for analy-
sis of XSDEO cyclodextrin inclusion complex [48].
Applications of essential oil‑inclusion complexes
Food applications
In food science, essential oils have been largely used as a
condiment, flavoring agent and food grade preservative, in
novel packaging system. Cinnamon oil is widely employed
in cooking, as a condiment and flavor for desserts, candies,
chocolate, tea and liquor [32]. Other essential oils/ essential
oil components employed in food manufacturing include
lavender oil [33], lemon oil, peppermint oil (−) linalool,
d-limonene, sweet orange, essential oil of Mentha × villosa
Hudson and essential oil of S. sclarea L. which are used for
their flavoring properties as food additives (Table 1). Flavor
is a mixture of aroma of compound or fragrant essential
oil, added to enhance the original taste or aroma of a food,
without providing aroma or taste of its own [94]. Mostly,
the components of flavor are highly volatile liquids, which
may undergo volatilization or oxidation on exposure to
heat or light. Some essential oils such cinnamon leaf oil
and garlic oil, estragole and eugenol have been used as food
preservatives. Due to antimicrobial, antifungal, antiviral
and insecticidal properties, these essential oils, via (−) lin-
alool, cinnamon leaf oil and garlic oil, estragole have also
been approved by FDA (Food Drug and Administration) as
GRAS substances to be employed as food additives [95].
Besides these applications, the essential oils also possess a
number of medicinal uses, which add to their utility as food
ingredients.
The direct utilization of essential oils is limited due to
their poor stability (in presence of oxygen, heat and light),
solubility and bioavailability. Therefore, these oils have been
encapsulated into cyclodextrin complexes. Encapsulation not
only enhances solubility, stability and bioavailability but also
masks the objectionable taste and odor of volatile oxidation
products. In addition, such complexes result in controlled
release products. Furthermore, α, β, γ-cyclodextrin have also
been approved as additive in the European Union, for food
industry, as well as granted GRAS status [34]. Cyclodextrins
are known to have adequate thermal and chemical stability
[96]. β-amylases and glucoamylases do not hydrolyze cyclo-
dextrins. However, they are more suspectible to α-amylases
(salivary and pancreatic amylase) or by hydrolysis. Another
important attribute is that cyclodextrins possess negligible
cytotoxic effects.
Ciobanu et al. evaluated complexation efficiency of α, β
or γ cyclodextrin for 13 volatile flavor compounds: α-pinene,
eucalyptol, linalool, limonene, camphene, β-pinene,
13

ρ-cymene, myrcene, menthol, menthone, trans-anethole,
camphor and pulegone, and observed β-cyclodextrin to be
the most versatile for studing guest molecules [34]. Petro-
vic et al., encapsulated cinnamon oils with β-cyclodextrin
successfully, using co-precipitation method and, reported
15:85 as the optimum ratio of essential oil to β-cyclodextrin
for commercial acceptance of this complex. Further, it
was reported that encapsulated essential oil maintained
its pharmacological properties and organoleptic behavior
[32]. Kfoury et al. complexed estragole from basil and tar-
ragon essential oils with cyclodextrin and demonstrated that
molecular inclusion allowed the controlled release of the
estragole [37]. Ayala-Zavala et al., encapsulated cinnamon
leaf and garlic oils using β-cyclodextrin and, the resultant
complexes illustrated good anti-fungal activity, which can
have application in the food technology [29].
Pharmaceutical and cosmetic applications
Cyclodextrins are equipped with the ability to temporarily
modify undesirable physicochemical properties of guest
moieties. Therefore, they have been explored for entrapment
of essential oils in molecular inclusion complexes [20]. In
this context, cyclodextrins have been reported to enhance
stability, improve the solubility characteristics and oral bio-
availability of essential oils. Table 2 summarizes essential
oil–cyclodextrin inclusion complexes reported for pharma-
ceutical applications.
Stability plays an important role in pharmaceutical for-
mulations, particularly for chemically labile moieties, like
essential oils or their bio-actives. It has been well empha-
sized that cyclodextrins help in improvement of stability of
various labile drugs against hydrolysis, dehydration, pho-
todecomposition and oxidation, leading to better shelf life
of these drugs [14]. Due to molecular shield, cyclodextrin
complexes protect the labile guests against the degradation
processes [97]. Tian and his research group demonstrated
that S. sclarea L. essential oil–β-cyclodextrin complexes led
to protection of this essential oil inside the cyclodextrin cavi-
ties, expanding its applications [42].
Essential oils or their components generally possess poor
water solubility hampering their bioavailability. This limits
their applications in pharmaceutical field. Xiang–Fu–Si–Wu
Decoction essential oil (XFSWD) is poorly water soluble,
volatile and possesses pungent smell [68, 98, 99]. The oil is
also reported to be sensitive to oxygen, humidity, light and
high temperature. This essential oil shows decreased bioa-
vailability due to poor water solubility. Additionally, pungent
smell is a reason for the poor compliance for oral administra-
tion [48]. Similar oral bioavailability enhancement results
were obtained with β-caryophyllene/β-cyclodextrin inclu-
sion complexes by Liu et al. [47].
13
J Incl Phenom Macrocycl Chem
To be pharmacological active, the drugs not only need
to possess some aqueous solubility but also, most drugs
require to be lipophilic for their permeation through bio-
logical membranes. Poorly soluble drugs are classified
as class II (poorly soluble/highly permeable) or class IV
(poorly soluble/poorly permeable) drugs, as per the Biop-
harmaceutics Classification System [100]. Hence, formu-
lations of such drugs that increase their apparent water
solubility, without affecting lipophilicity, play a major role
in enhancing their clinical applications [71]. Cyclodextrins
have been reported for apparent drug solubility enhance-
ment through inclusion complexation, as a hydrophilic
carrier. The capacity as solubilizer also depends on type
of cyclodextrin used. For example, eugenol, major compo-
nent of clove oil, possesses a wide spectrum of biological
activities. However, the limitations such as poor solubil-
ity, pungent taste, degradation and irritation characteristics
towards the mucosa, make it unsuitable for use. As per
Garg et al., inclusion complex of eugenol with hydroxy
propyl-cyclodextrin, prepared by lyophilization method
retained oxidation potential and resulted in better solubil-
ity [45].
In cosmetic industry, essential oils can be employed
as fragrance compounds in soaps, perfumes, colognes,
shampoos and skin lotions. However, the above-men-
tioned limitations limit their use in cosmetic products as
well. Additionally, due to reactive functionalities such as
ketone, aldehyde and terpenes, degradation of fragrance
compounds occurs, leading to changes in their sensory
features, and eventually forming allergenic products
[101–103]. Hence, encapsulation as cyclodextrin com-
plexes may be adopted to improve the stability of many
fragrance molecules.
Therapeutic applications
As mentioned earlier, essential oils have been known to
possess diverse biological activities. They are potential
sources of naturally derived compounds with several func-
tions, such as anti-fungal, anti-microbial, antioxidant,
anticonvulsant, analgesics, anticancer and antinociceptive
effect. Additionally, some bio-actives like citronella and
geraniol possess mosquito repellent activity. Encapsulation
of these oils and their components improves their phys-
icochemical properties, without affecting the biological
efficacy.
As reported in Table 3, essential oil–cyclodextrin com-
plexes have been investigated for therapeutic effectiveness
by various research groups. Tao et al. encapsulated thymol
and thyme oil in β-cyclodextrin inclusion complexes and
investigated their physicochemical characteristics, along
with anti-microbial activity against food borne bacteria [59].
J Incl Phenom Macrocycl Chem
Conclusion
Developments in drug design technology over the years
have yielded numerous active moieties, whose undesirable
properties, such as chemical or physical unstability, irrita-
tion, toxicity, poor solubility/dissolution and bioavailability
have been a deterrent factor in their clinical utility. Amongst
the various strategies to overcome such problems, use of
cyclodextrins has been widely investigated and even found
promising in alleviating drug delivery issues. This is fur-
ther supported by the huge number of publications, patents
and approved products. Although, carbohydrates in various
forms have been used in food products, their degradation
products, cyclodextrins, since their inception, opened new
avenues not only for the food industry but for pharmaceuti-
cal and cosmetic industry, as well. With the use of cyclo-
dextrins and their modified forms, it is possible to entrap
active agents at a molecular level in cyclodextrin inclusion
complexes.
Essential oils are known to possess properties which
make them worthy of application in number of fields, includ-
ing food, pharmaceuticals and cosmetics. For maximum uti-
lization of the potential benefits of essential oils, cyclodex-
trin complexes represent a very attractive tool to overcome
their limitations and facilitate ease of handling.
Despite the vast literature published in context of essen-
tial oil inclusion complexes, there are certain issues which
demand the attention of research community and further
investigation. The most important objective of fabricating
inclusion complexes is to attain maximum inclusion effi-
ciency. In this concern, type of cyclodextrin used, guest–host
ratio and method of preparation constitute the most sig-
nificant parameters, which need to be carefully optimized.
Notwithstanding the research carried out by certain inves-
tigators, the studies should be more systemically designed,
focusing on optimization of the above-mentioned param-
eters. Moreover, one should not overlook the fact that differ-
ent essential oils may have different retention and inclusion
efficiency in cyclodextrin complexes.
Though variety of techniques have been reported for
the preparation of essential oil cyclodextrin complexes,
the peculiar physical and chemical nature of essential oils
calls for comparative studies, with the view to find the most
suitable preparation process. Furthermore, the available lit-
erature suggests that β-cyclodextrins are the most explored
amongst all the forms of cyclodextrins, while the other
forms have received little attention. It is further emphasized
that the investigations performed in this domain should be
taken a step ahead of formulation development and charac-
terization, so as to validate the claimed activities. With the
advent of more sophisticated analytical tools and use of in
silico approaches, the understanding of structural features of
essential oil inclusion complexes can be further bolstered.
Compliance with ethical standards 
Conflict of interest  All of the authors have no conflict of interest to
declare.
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